| Method |
Sitting-drop vapour diffusion for initial crystal screening and hanging-drop vapour diffusion for crystal optimization |
| Plate type |
96-well plates for initial crystal screening and 24-well plates (Hampton Research HR3-107) for crystal optimization |
| Temperature (°C) |
4 |
| Protein concentration (mg ml−1) |
12 |
| Buffer composition of protein solution |
50 mM Tris–HCl pH 7.5, 100 mM sodium chloride, 20 mM β-mercaptoethanol |
| Composition of reservoir solution |
0.1 M MES monohydrate pH 6.5, 1.6 M magnesium sulfate heptahydrate |
| Volume and ratio of drop |
1.0 µl for initial crystal screening and 2.0 µl for optimization; 1:1 and 2:1 protein:reservoir solution |
| Volume of reservoir |
50 µl for initial crystal screening and 1.0 ml for optimization |
| Composition of the cryoprotectant |
0.1 M MES monohydrate pH 6.5, 1.6 M magnesium sulfate heptahydrate, 30% ethylene glycol |
| Drop setting |
Drop setting was performed manually using a micropipette. Crystallization drops were prepared by mixing protein and reservoir solutions at the specified ratios of 1:1 (screening) and 2:1 (optimization). |
| Seeding |
No |
